番茄褪绿病毒与番茄黄化曲叶病毒复合侵染的分子鉴定

Molecular Identification on Mixed Infections of Tomato chlorosis virus and Tomato yellow leaf curl virus

2013年秋季,在山东寿光设施番茄植株上采集到叶片脉间褪绿黄化、叶片边缘卷曲、叶片皱缩,后期叶片变厚、变脆,并伴随着植株矮化的疑似番茄褪绿病毒和番茄黄化曲叶病毒复合侵染的样本。分别利用番茄褪绿病毒(Tomato chlorosis virus,To CV)外壳蛋白基因(CP)序列引物CP-F/CP-R和番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)特异引物TY1-R/TY1-F、TY2-R/TY2-F、TY3-R/TY3-F对其进行检测、扩增,分别得到774bp(Gen Bank登录号KC709510)和2781bp(Gen Bank登录号KJ546418)的核苷酸序列。经序列测序后表明,KC709510与Gen Bank已登录的番茄褪绿病毒To CV-BJ(KC311375)分离物相似性为99.6%,KJ546418与Gen Bank已登录的番茄黄化曲叶病毒TYLCV-SDWF-L7(KC999850)分离物相似性为99.6%。

In the autumn of 2013, leaves of greenhouse-grown tomato plants showed symptoms of interveinal chlorosis, and together with stunted, marginal curling and crumpling. As the disease progressed, leaves become thick and crisp, similar to symptoms induced by Tomato chlorosis virus （ToCV） and Tomato yellow leaf curl virus （TYLCV） were collected from greenhouse-grown tomato plants in Shouguang, Shandong Province. The specific primer pairs of CP-F/CP-R specific to ToCV and TY1-R/TY1-F,TY2-R/TY2-F,TY3-R/TY3-F specific to TYLCV were used to detect ToCV and TYLCV, respectively. 774 bp （GenBank accession number： KC709510） and 2 781 bp （GenBank accession number： KJ546418） specific fragments were amplified from the same symptomatic samples. Sequence analysis of the amplified sequence of KC709510 showed 99.6% nucleotide sequence identity with ToCV-BJ （KC311375） isolates and the sequence of KJ546418 showed 99.6% nucleotide sequence identity with TYLCV-SDWF-L7 （KC999850） isolates registered in the GenBank.